Fluorescent method for detection of cleaved collagens using O-phthaldialdehyde (OPA)

Go, Katrina V.; Horikawa, Yousuke T.; Garcia, Ricardo Alexandrino; Villarreal, Francisco

doi:10.1016/j.jbbm.2007.05.004

Cited by 22 publications

(13 citation statements)

References 9 publications

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“…The release of amino acids by the proteolysis of proteins forming wheat flour and coffee fibre was measured by the OPA assay according to Go et al [ 15 ]. OPA reagent was freshly prepared by dissolving 10 mg OPA in 250 μL ethanol (95%, v/v ), 9.8 mL phosphate buffered saline solution PBS (10 mM, pH 7.4) and 20 μL β-mercaptoethanol.…”

Section: Methodsmentioning

confidence: 99%

Biscuits with No Added Sugar Containing Stevia, Coffee Fibre and Fructooligosaccharides Modifies α-Glucosidase Activity and the Release of GLP-1 from HuTu-80 Cells and Serotonin from Caco-2 Cells after In Vitro Digestion

et al. 2017

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This study assessed the in vitro effects of the bioaccessible food components released during the simulated human digestion of a coffee fibre-containing biscuit (CFB) on α-glucosidase activity, antioxidant capacity and satiety hormones. Digest of CFB presented a significantly (p < 0.05) lower amount of sugar (68.6 mg/g) and a higher antioxidant capacity (15.1 mg chlorogenic acid eq./g) than that of a sucrose-containing biscuit (SCB). The CFB significantly reduced (p < 0.05) α-glucosidase activity (IC50 = 3.3 mg/mL) compared to the SCB (IC50 = 6.2 mg/mL). Serotonin and glucagon-like peptide-1 (GLP-1) release by differentiated Caco-2 and HuTu-80 cells, respectively, was stimulated by the CFB (355% at a concentration of 0.5 mg/mL and 278% at a concentration of 0.05 mg/mL) to the same order of magnitude as those of the SCB. To summarize, the CFB was demonstrated to reduce monosaccharide bioaccessibility, to inhibit a diabetes-related digestive enzyme, and to improve the release of satiety hormones.

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Section: Methodsmentioning

confidence: 99%

Biscuits with No Added Sugar Containing Stevia, Coffee Fibre and Fructooligosaccharides Modifies α-Glucosidase Activity and the Release of GLP-1 from HuTu-80 Cells and Serotonin from Caco-2 Cells after In Vitro Digestion

et al. 2017

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“…Left ventricle samples from perfused hearts were used. Myocardial collagen was extracted using sequential ethanol/salt precipitations as described previously 33. Collagen samples isolated from rat heart ventricles (100 mg each) were used for o -phthaldialdehyde conjugation to exposed amino termini (“opalation”) as described previously 33.…”

Section: Methodsmentioning

confidence: 99%

“…Myocardial collagen was extracted using sequential ethanol/salt precipitations as described previously 33. Collagen samples isolated from rat heart ventricles (100 mg each) were used for o -phthaldialdehyde conjugation to exposed amino termini (“opalation”) as described previously 33. Collagen content was calculated under the assumption that the average hydroxyproline content of collagen is 10%, and thus 1 μmol of hydroxyproline is equivalent to 1 mg of collagen.…”

Section: Methodsmentioning

confidence: 99%

Effects of Novel Semiselective Matrix Metalloproteinase Inhibitors on Ex Vivo Cardiac Structure-Function

Romero-Perez

Agrawal²,

Jacobsen³

et al. 2009

Journal of Cardiovascular Pharmacology

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The purpose of this study was to evaluate the ability of novel semiselective matrix metalloproteinase inhibitors (MMPI) to protect myocardial structure-function in the setting of ischemia-reperfusion injury. For this purpose, an isolated rat model of myocardial stunning and infarction was used. Isolated hearts were subjected to 20-30 minutes of global no-flow ischemia and 30-minute reperfusion. Myocardial performance was assessed as the product of the heart rate and left ventricular developed pressure (rate-pressure product, RPP). Coronary flow rates, ventricular weights, indicators of muscle (troponin I), and fibrillar collagen damage (collagen opalation) were measured. Four MMPI were tested: 2 non-hydroxamate, semiselective inhibitors (PY-2 and 1,2-HOPO-2) and 2 broad-spectrum inhibitors (PD166793 and CGS27023A). The non-hydroxamate, semiselective inhibitors were shown to be nontoxic in cocultures of cardiac cells. Results indicate that semiselective inhibitors (in particular 1,2-HOPO-2) yield improved cardiac performance (~23% higher RPP vs. controls) and coronary flow rates (~22%), reducing muscle (~25%) and fibrillar collagen damage (~60%). Evidence suggests the involvement of matrix metalloproteinase-2 in these actions. Interestingly, broad-spectrum inhibitors only show modest improvement (~8% higher RPP vs. controls) without affecting the other measured parameters. In conclusion, semiselective MMPI can act as cardioprotectors in isolated perfused rat hearts. Protection is observed in all structural components of the myocardium translating into improved contractile function. Based on these findings, non-hydroxamate, semiselective MMPI warrant further studies as to their ability to protect ischemic myocardium in the in vivo setting.

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“…Over the years, fluorescent labeling reagents offer advantages in protein assays . Protamine quantification in pharmaceutics, detection of cleaved collagens, lysine determination in wheat gliadin proteins, and protein content of aluminum‐based adjuvants are the assays that are performed by OPA reagents in recent years.…”

Section: Introductionmentioning

confidence: 99%

Rapid quantification of total protein with surface‐enhanced Raman spectroscopy using o‐phthalaldehyde

Eryılmaz

Zengi̇n

Boyaci

et al. 2017

J Raman Spectroscopy

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In this study, surface-enhanced Raman spectroscopy (SERS) based quantification method for the total protein using o-phthalaldehyde is reported for the first time. For this purpose, o-phthalaldehyde was chosen to form a complex with protein, and SERS signal was observed in the presence of gold nanoparticles. A calibration curve was obtained by plotting the intensity of the SERS signal at 727 cm À1 versus the concentration of protein standard (bovine serum albumin, BSA). Thus, the correlation was found to be linear within the range of 0.054-0.72 mg/ml of BSA, and the limit of detection was determined to be 0.08 mg/ml. All tests were carried out using a portable Raman instrument with an analysis time of 5 min. In addition, the ability of the developed method to quantify total protein in milk samples was investigated, and the obtained results were compared to the conventional ultraviolet methods.

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Fluorescent method for detection of cleaved collagens using O-phthaldialdehyde (OPA)

Cited by 22 publications

References 9 publications

Biscuits with No Added Sugar Containing Stevia, Coffee Fibre and Fructooligosaccharides Modifies α-Glucosidase Activity and the Release of GLP-1 from HuTu-80 Cells and Serotonin from Caco-2 Cells after In Vitro Digestion

Biscuits with No Added Sugar Containing Stevia, Coffee Fibre and Fructooligosaccharides Modifies α-Glucosidase Activity and the Release of GLP-1 from HuTu-80 Cells and Serotonin from Caco-2 Cells after In Vitro Digestion

Effects of Novel Semiselective Matrix Metalloproteinase Inhibitors on Ex Vivo Cardiac Structure-Function

Rapid quantification of total protein with surface‐enhanced Raman spectroscopy using o‐phthalaldehyde

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